The pharmaceutical industry has focused on the identification of small molecule compounds that modulate (e.g., block, reduce, or enhance) specific steps in biological pathways. Lead Discovery, the process of identifying, cloning, expressing, and running a high throughput screen (HTS) for the purpose of identifying lead chemical matter as a starting point for development of therapeutic drugs plays a key role in drug discovery. Historically, this has meant a focus on those targets that are considered readily amenable to HTS. A test or assay for HTS must measure a relevant activity of the target, produce a robust signal for that activity, be readily assembled from commercial and/or in-house prepared reagents, and entail the minimum possible sequence of physical steps involving addition or sampling of the test reagents/mixture. In this way, a large volume of samples may be evaluated in a short period of time, and enables for automation of steps.
In general, because of time and labor constraints for assay methods, lipid metabolism proteins have not been assayed by HTS. However, because of the diverse array of chemical matter within HTS compound files having pharmacological potential for lipid metabolism targets, there exists a distinct need to prosecute these targets through HTS.
Lipids play an indispensable role in cell structure, metabolism, and cell signaling. For example, fatty acids and triacylglycerols are the major storehouse of metabolic energy. Enzymes that digest, oxidize, and synthesize these energy storage forms are potential therapeutic intervention points for drug discovery, for example, in the areas of cancer, obesity, cardiovascular, and diabetes research. In addition to their metabolic role, phospholipids play key roles in signal transduction pathways, and biological macromolecules that control phospholipid metabolism are also potential drug discovery targets. Traditional assay methods for lipid metabolism targets, including thin layer chromatography (TLC) and high-pressure liquid chromatography (HPLC), are labor- and time-intensive. Thus, there is a need in the pharmaceutical industry to develop methods to screen lipid metabolism targets that are efficient and cost-effective.